DNA Replication and Manipulation Flashcards

Question

Telomerase

Answer 1

Solution to end replication problem GERM CELLS ONLY (healthy) in somatic cells, teomerase turned on leads to cancer

Answer 2

* Telomerase enzyme contains an RNA template that allows the template strand to be lengthened by telomere repeats. * Germ line cells are “immortal”

Answer 3

long stretch of repetitive DNA at end of somatic cells to deal with end replication problem so you lose repeat, not important genes, with division

Answer 4

polymerase from themoatically stable organism (one that lives in hot environments) targeted region of DNA to be replicated/amplified into as many copues as desired

Answer 5

Swimming Pool ex (SI) • DNA samples are inserted into wells at one edge of gel • Pass electricity through (also DNA matrix and liquid buffer) • DNA migrates to positive charge at opposite end of chamber • Passes through substrate depending on size • Small pieces pass rapidly • Numbers on pictures are wells Bigger stay at negative, little go to positive

Answer 6

restriction enzymes used to cut up whole DNA separate using Gel electrophinesis pass DNA from gel to filter paper look for where fragment of interest migrated probe filter paper with radiolabeled matching gene probe expose film, see where bands migrated can use flourencent nucleotides instead of raido

Answer 7

Denaturation Annealing Extension

Answer 8

Heat up strands and they come apart | Unwinding

Answer 9

When cooled, two primers provided to anneal to their complementary sequences on the DNA template strand o need temp not too low, or DNA will zip back together

Answer 10

Synth new DNA strands | Extend primers in a 5’ to 3’ direction

Answer 11

o Template DNA o DNA Polymerase (TAQ – polymerase from Thermus aquaticus) o The four deozynucleoside triphosphates (A, T, G, or C) o Two primers (short stretches of DNA).

Answer 12

o Target specific segement of DNA • 1. Template duplex often longer than amplified region o provide primers that match on either side of DNA o let amplification occur o end up with 2 copies: parental and new o Go through again, end up with 4 copies o After n cycles of amplification there are 2^n copies of the template sequences. o Shorter copies in each subsequent round

Answer 13

Cut DNA at specific sequences allows: • pieces from the same or different organisms to be brought together in recombinant DNA technology • or can cut large genomes into small pieces for further analysis

Answer 14

Even on either side of cut

Answer 15

Overhang on either side of cut

Answer 16

Don't have 3' hydorxyl group, just hydrogen, so others cant add on

Answer 17

o Run fragments out on gel o Read gels looking at readout o Florecnese peak tells us what last nucleotide was o Read one fragment at a time, put all together, find sequence of daughter strand

Answer 18

``` usually e. coli used Transformed Take up DNA from surrounding environment "Heat shock" makes them stressed and suck up plasmid. Has its genome and the one you gave it. ```

Answer 19

The one we add in PCR replicate it (double stranded from us sometimes) Ligate to vector with ligase of sticky ends

Answer 20

circular pieces of DNA taken up. From e. coli. Cut it Ligate with sticky ends to Donor

Answer 21

Gene gun engineer viruses to insert into eukaryotic genomes maybe review this

Answer 22

Don't know template, so make daughter to ID parent

Answer 23

to replicate the gene you want.

Answer 24

make new type of e. coli cells with its genome and recombiant plasmid Make lots of it to replicate the gene of interest

Answer 25

Pass DNA from gel electro to filter paper want to know where DNA fragment of interest is Probe filter paper (probe matches gene) visualize by exposing film